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PubMed1990

Hydatidosis in Belgium: analysis of larval Echinococcus granulosus by SDS-PAGE and western blotting.

Echinococcosis/hydatidosis is characterized by the existence of various strains of the parasite Echinococcus granulosus. In order to study the degree of variability in Belgium, protoscoleces (PS) were isolated from infected intermediate hosts (horse, cattle, pig, goat and man). Homogenate supernatants of these PS were compared by SDS-polyacrylamide gel electrophoresis and western blotting: 15 to 26 predominantly silver staining bands (depending on the source of the parasites), were selected to study existing homologies among these possible strains. Antisera raised against PS homogenates of horse, cattle, pig and goat origin, were used to compare antigenicity by western blotting. The results indicate a closer similarity between E. granulosus of horse, goat and pig origin whereas E. granulosus derived from cattle showed less homology with any of the former. Results are discussed against the background of the occurrence of E. granulosus in Belgium.

Animals↗
PubMed1990

[Uptake and distribution of [3H]praziquantel in Echinococcus granulosus cysts].

[3H]Praziquantel was absorbed rapidly by the cysts in mice infected with metacestodes of Echinococcus granulosus and treated with ig [3H]praziquantel 400 mg/kg (containing 3H 1.48 MBq). Within 24 h after treatment, 3H content in the ectocyst was 27.7-54.9% higher than that in cyst wall. The peak concentration of 3H in cyst wall and cyst fluid were 11.7-15.3% of those found in plasma. However, when the cysts were exposed to [3H]praziquantel 20 micrograms/ml (containing 3H 3.7 kBq), the 3H radioactivity in the cyst wall and cyst fluid reached equilibrium within 30 min-48 h after medication, corresponding to 45.5-63.3% of 3H radioactivity in the medium. Autoradiography showed that [3H]praziquantel entered into the germinal layer through ectocyst and laminated layer. The damage to germinal layer was related closely to deposition of the drug.

Animals↗
PubMedDate not supplied

[Experimental imaginal Echinococcus multilocularis infestation of golden hamsters].

The imago stage of Echinococcus multilocularis development (prior to maturation of parasite oocysts) has been described in the intestine of golden hamsters infected by parenteral penetration of an Alveococcus agent and given hydrocortisone subcutaneously for 3 days before the infection and on the day of the infection, mean total drug dose being about 250 mg/kg. The number of helminths developed on days 8-35 of postinvasion ranged from 432 to 1863, their length was 1.0 to 4.0 mm and the number of segments did not exceed 2. Oocyst maturation was noted on day 20 of postinvasion. High oocyst invasiveness was confirmed by a biological test on 32 cotton rats aged 1-1.5 months who were administered parenterally 2-3 helminths containing 86 to 284 oocysts. All the rats died of severe hepatic alveococcosis on days 24-43 of postinfection.

Animals↗
PubMed1990

Effects of ethyl N-N-benzyl-methyl-oxamate in Meriones unguiculatus infected with Echinococcus multilocularis metacestodes. Biochemical and ultrastructural observations.

Ethyl N-N-benzyl-methyl-oxamate is a N-substituted derivative of the oxamic acid, well-known as inhibitor of the lactate dehydrogenase activity. The biochemical and ultrastructural effects of this drug in Echinococcus multilocularis metacestodes and the repercussions in the livers of receptive host, Meriones unguiculatus, were investigated. This compound decreased the specific activity of the lactate dehydrogenase in the metacestodes and the host liver by 81 and 86.8%, respectively. This N-methyl compound has shown an important repercussion on the lactate dehydrogenase activity due to its greater lipophilicity and thus allows a better penetration of the molecule. In the host liver, a fall of the alkaline and acid phosphatases activity was observed. The glucose and glycogen concentrations were also decreased. The ultrastructural study provided an alteration of the tegument of the metacestodes and damages of the muscular system. The parenchyma was disorganized. In conclusion, these biochemical and ultrastructural data obtained with ethyl-N-N-benzyl-methyl oxamate in E. multilocularis metacestodes show clearly the importance of the lactate dehydrogenase activity in the metacestodes.

Acid Phosphatase↗
PubMed1989

[Characterization of immunogenic proteins from Echinococcus granulosus with the immunoblot technique].

The serological response to Echinococcus granulosus was evaluated by immunoblotting in 20 patients with hydatid disease. In non-reducing conditions, the highest reactivity corresponded to 80, 66 and 56 kd proteins, while in reducing conditions only one 36 kd protein was detected. A low molecular weight component which moved with the migration front of gels was also detected. In reducing conditions the activity was less marked. Immunoblotting permits the identification of specific immunogenic proteins and the improvement of the serologic diagnosis of hydatid disease, as it eliminates the problem of low titer nonspecific reactions with serological tests using complex antigens.

Animals↗
PubMed1989

[Effect of praziquantel and albendazole on ultrastructure of protoscolex of Echinococcus granulosus].

When protoscoleces of Echinococcus granulosus, maintained in 20% calf serum-RPMI 1640 were exposed to praziquantel 1 microgram/ml for 1-48 h, severe damages to the ultrastructure of protoscoleces were observed. The main lesions of the tegument included indistinct of the matrix, vacuolization and peeling, while vacuolization of perinuclear cytoplasma in tegumental cells, focus lysis in muscle bundles, and destruction in collection ducts and flame cells were also seen. The results suggest that the damages to the tegument, collection ducts and flame cells may interfere with the nutrition and defence functions of protoscoleces, and destroy the osmoregulatory system, which may be the major causes of death in protoscoleces during treatment with praziquantel. When protoscoleces were exposed to albendazole 20 micrograms/ml for 24-72 h, no apparent damage to the tegument was detected, except that some network-like structures appeared in the cytoplasma of tegument and parenchyma cells.

Albendazole↗
PubMed1989

[Cell line of hybridoma secreting monoclonal antibodies against Echinococcus granulosus antigens].

In order to produce specific monoclonal antibodies for immunodiagnosis and immunoprophylaxis in hydatid disease, a monoclonal antibody cell line NIA2 secreting specific antibodies was established, through fusion of myeloma cells SP2/0 and spleen cells of BALB/c mice immunized with hydatid fluid from a local (Ningxia) patient and repeated screenings and clonings. The specificity was assayed via ELISA, immunoprecipitating and ultramicroscopy. Antibody titer of 1:2,560 was obtained in the supernatant of the culture medium by ELISA and a high titer of 1:163,840 in seeded ascitic fluid of mice of the same isolate. Immunoglobulin subclass of the NIA2 secreting antibody was determined to be IgG1 which showed no cross reaction to Cysticercus cellulosae. Cloned cell lines from the hybridomas were found consistent in secreting anti-Echinococcus granulosus antibody through serial passage in tissue culture for ten months.

Animals↗
PubMed1989

Activation of normal murine B cells by Echinococcus granulosus.

Echinococcus granulosus protoscolex (PSC) infection of BALB/c mice led, after 4 days, to raised numbers of cells forming plaques with trinitrophenyl-treated sheep red cells and bromelain-treated mouse red cells. The findings were similar in athymic and euthymic CBA mice. Activation of B cells was accompanied by secretion of immunoglobulin, as indicated by the reverse plaque technique. In addition, co-culture of PSC with the 7OZ/3 pre-B-cell led to the induction of differentiation, resulting in the expression of surface immunoglobulin (Ig). It is concluded that E. granulosus is a polyclonal activator of B cells inducing both transformation and differentiation, and that the effect is thymus-independent.

Animals↗
PubMed1989

[Production and characterization of monoclonal antibodies to Echinococcus multilocularis].

Spleen cells from BALB/c mice immunized against Echinococcus multilocularis were fused with mouse myeloma cells (P3X6.5.3.) in the presence of polyethylene glycol (PEG 1000), and the hybridomas were obtained. Of the 85 hybridomas, 23 were found to produce antibodies. Of the 23 cell lines, 13 hybridomas which produced a high titer of antibodies were cultured for cloning the cells by the limiting dilution method. Then 5 monoclonal antibodies were obtained. The immunoglobulin class of the monoclonal antibodies were IgM in one, IgG in 4. IgG monoclonal antibodies were studied by the ABC immunostain method. The germinal layer, brood capsules and protoscolices were stained. Germinal layer was especially stained by 4 monoclonal antibodies. However sections of liver, kidney, spleen and other parasites; anisakis, ascariasis lumbricoides, entamoeba histolytica and schistosoma japonicum were not stained with those monoclonal antibodies.

Animals↗
PubMed1989

[Immunological humoral response in mice experimentally infected with Echinococcus granulosus, preliminary result of kinetic observation].

5 serological methods included latex agglutination test (LA), enzyme-linked immunoelectrotransfer blot (EITB), enzyme-linked immunosorbent assay (ELISA), dot-enzyme-linked immunosorbent assay (Dot-ELISA) and immunofluorescence antibody test (IFA) were used to trace the kinetic changes of the humoral response in NIH mice experimentally infected with protoscolex of Echinococcus granulosus. Results showed that Dot-ELISA and IFA were able to detect antibodies 2 wk after infection while ELISA, EITB and LA gave positive reactions 4 wk, 2 mos and 3 mos after infection respectively. Therefore Dot-ELISA and IFA were considered to be the most sensitive ones among the 5 methods used. All infected mice gave positive reactions at 8 and 12 mos but none of the normal control mice showed any reaction throughout the period of observation (Fig. 1).

Animals↗
PubMed1989

Orbital hydatid cyst of Echinococcus oligarthrus in a human in Venezuela.

This study reports the first known case of human hydatid disease caused by the larval stage of Echinococcus oligarthrus. The patient, a native Venezuelan female, presented a single cyst localized intraorbitally behind the left eye (retroocular), which was discovered by computed tomography. The cyst was removed by surgery, and after parasitological studies it was identified as an E. oligarthrus hydatid cyst. This is also the first case of intraorbital hydatid cyst in humans in Venezuela.

Animals↗
PubMed1989

The dispersion of Echinococcus granulosus in the intestine of dogs.

We studied the dispersion of adult Echinococcus granulosus in the intestine of experimentally infected dogs at 2 scales of habitat use. On a coarse scale, worms were found most frequently in the anterior third of the small intestine. On a fine scale, clumps or aggregations, typically of 4-5 worms in an area of 12 mm2, occurred throughout the anterior two-thirds of the intestine. The most likely proximate cause of aggregative behavior is attraction between individual worms. There are at least 2 equally plausible ultimate causes of the behavior: to enhance cross-fertilization and to improve the quality of the environment. Restriction of worms to the anterior small intestine may be a consequence of aggregative behavior on a finer scale or a response to different proximate and ultimate factors.

Animals↗
PubMed1989

[Echinococcus multilocularis in a swamp beaver (Myocaster coypus)].

Larval Echinococcus multilocularis was found in a nutria (Myocastor coypus) on a fur farm situated south of the Thuringian Forest near the border the Federal Republic of Germany. This finding is of epidemiological importance. Investigations of 67 red foxes (Vulpes vulpes) originating from this area and the whole Erfurt Region (Thuringia) did not reveal the presence of adult E. multilocularis.

Animals↗
PubMed1988

[Characterization of surface and intracellular glycoconjugates in larval forms of Echinococcus multilocularis using lectins].

Carbohydrate moieties have been detected with nine fluorescent lectins in Echinococcus multilocularis cysts, developed in liver and lung of infested jirds. Tegument and glycocalyx, and two types of medullary cells were found to selectively and strongly bind lectins which are specifically adsorbed by N acetyl glucosamine, N acetyl galactosamine, galactose or mannose. The significance of these features in the host-parasite interaction are discussed.

Animals↗
PubMed1987

[In vivo inhibition by ethyloxanilates of alkaline phosphatases of the cestode Echinococcus multilocularis].

Alkaline phosphatases (E.C.3.1.3.1), membranous enzymes of the cestode Echinococcus multilocularis have been studied in the parasite and in the experimental host liver. Synthetic inhibitors interaction with metacestode alkaline phosphatases is reported. In regard to the alkaline phosphatases inhibition, the ethyloxanilate 2 is more efficient in the cestode itself than in the host liver.

Alkaline Phosphatase↗
PubMed1988

Studies on the effect of short term, high dose praziquantel treatment against protoscoleces of ovine and equine Echinococcus granulosus within the cyst, and in vitro.

Protoscoleces of Echinococcus granulosus were subjected to high concentrations of praziquantel (10-1000 micrograms/ml medium or cyst fluid) for a short, 10 min duration in vitro or in situ within cysts. Drug efficacy was then monitored either by in vitro culture in drug-free medium and/or by passage into rodents. The in vitro test of effectiveness suggested that ovine-derived protoscoleces were more sensitive than those of equine origin, and that the in vitro method of treatment was more successful than that performed in situ. However, the subsequent passage of treated protoscoleces, whether immediately after in situ or in vitro exposure or after culture in drug-free medium, indicated that the in situ administration was as effective as the in vitro treatment. Both methods reduced cyst burdens in rodents to 9-15% of control infections.

Animals↗
PubMed1988

Echinococcus strains in Europe: a review.

In European countries the following final/intermediate host assemblages of Echinococcus granulosus are known to occur: (a) dog/sheep; (b) dog/horse; (c) dog/cattle; (d) dog/pig, (e) dog/reindeer (= "northern form" of E. granulosus). The parasites involved in cycles a-d have been identified as distinct strains in various countries. Epidemiological aspects relevant to intraspecific variations and strain characteristics are described. In E. multilocularis there are some indications for intraspecific variability but it is yet unknown whether or not they are of epidemiological or other significance. There is clear evidence that E. multilocularis, the dog/sheep strain and the "northern form" of E. granulosus are infective for humans; the status of the other strains regarding infectivity to man is still unclear.

Animals↗
PubMed1986

Transformation induced by Echinococcus granulosus protoscoleces in unprimed murine spleen cells: identity and MHC restriction of participating cell types.

Transformation induced by Echinococcus granulosus in unprimed murine spleen cells in vitro was not prevented by rigorous T- or B-cell depletion. When naïve T- or B-cell populations were further depleted of macrophages, B-cell transformation was relatively unaffected whereas T-cell transformation was totally inhibited. Peritoneal macrophages from infected mice could present a mitogenic stimulus to purified, naïve T-cells, provided that the T-cells had identity in the I region of the H2 complex. K/D identity was neither necessary nor sufficient. It is concluded that transformation can be induced by E. granulosus in both T- and B-cells and that, at least in naïve T-cells, the stimulus can be an I region-restricted, macrophage-presented signal.

Animals↗